Isolation of Yeasts from Wild Flowers in Gyonggi-do Province and Jeju Island in Korea and the Production of Anti-gout Xanthine Oxidase Inhibitor

(1)

경기도와 제주도 야생화들로부터 효모의 분리, 동정 및 항통풍성 Xanthine oxidase 저해물질의 생산

현세희¹, 문혜연², 이향범², 김하근¹, 이종수¹*

1배재대학교바이오·의생명공학과

2전남대학교응용생물공학부

Received: September 6, 2013 / Revised: September 24, 2013 / Accepted: September 27, 2013

서 론

효모는유성생식과무성생식을하며진핵세포를갖고있 는진균류로서일반적으로자낭균류와일부담자균류에속 하는유포자효모와불완전균류에속하는무포자효모로크 게구분된다

^.

효모는대부분이

^GRAS

균주로서오래전부터 주류와장류등의발효식품제조에이용되고있고근래건 강식품소비의증가에따라효모에서항고혈압성안지오텐 신전환효소저해물질

^{[10, 12],}

항치매성β

-secretase

저해물 질

^[14]

과

acetylcholinesterase

저해물질

^[15],

혈전용해물질

[8],

혈관신생억제물질

^[9],

미백성

tyrosinase

저해물질

^[7],

정 미성리보핵산물질

^[16]

과같은다양한대체의약소재들이개 발되어보고되었다

^.

최근시판막걸리에서항고혈압활성이

우수한

Pichia burtonii

등이 분리 동정되었고

[21], killer toxin[17],

효소생산효모

^[18]

등이분리

^,

선별되어일부가산 업적으로이용되고있다

^.

또한

^,

효모의각종대사산물의생 성

^,

분비기작이규명되었고빠른생육과배양이용이한잇 점등이있어유전공학연구에서귀중한숙주로도이용되고 있다

^[11].

지금까지국내에서효모는주로전통발효식품이나누룩

^,

곡자

^,

발효제등에서분리

^,

동정되어이들의일부가산업적 으로이용되고있을뿐우리나라자연환경에분포하고있는 야생효모들을분리

^,

동정하고이들을산업적으로응용하는 연구는거의실시되지않았다

^.

따라서필자등은전국각지 의자연환경에서식하고있는야생효모들을분리

^,

동정하여 다양한효모균주들을확보하기위해전보

^[20]

에서대전과 충남일부지역에서서식하고있는야생화들로부터효모들 을분리

^,

동정하여보고하였고우리나라해안근접지역과금 산군

^,

안동시등의일부내륙지역야생화들로부터국내미기 록효모들을분리하여특성을보고하였다

^[5].

본연구에서는

Isolation of Yeasts from Wild Flowers in Gyonggi-do Province and Jeju Island in Korea and the Production of Anti-gout Xanthine Oxidase Inhibitor. Hyun, Se Hee

¹

, Hye Yeon Mun

²

, Hyang Burm Lee

²

, Ha-Kun Kim

¹

, and Jong-Soo Lee

¹

*.

1

Department of Biomedicinal Science and Bioechnology Paichai University, Daejeon 302-735, Korea,

²

Divison of Applied Bioscience & Biotechnology, Chonnam National University, Gwangju 500-757, Korea

Several yeasts were isolated from flowers found in Gyonggi-do Province and Jeju island in Korea. They were then identified by a comparison of their PCR-amplified D1/D2 regions of 26S rDNA, internal transcribed spacer 1 and 2 inclusive of 5.8S rDNA, using the BLAST database. A total of fifty four yeast strains were isolated from wild flowers in Gyonggi-do and the genus Pseudozyma was noted as being dominant. A total of thirty two strains were isolated from Songaksan and Seongsan-ilchul- bong in Jeju island and Sporobolomyces ruberrimus was seen to be dominant. The anti-gout xanthine oxidase inhibitory activi- ties of the culture broths and cell-free extracts from eighty six yeast strains were then determined. The cell-free extracts of Pseudozyma hubeiensis 228-S-1 exhibited the highest xanthine oxidase inhibitory activity of 19.6%. The XOD inhibitor was also maximally produced when Pseudozyma hubeiensis 228-S-1 was cultured at 30

^o

C for 36h in YEPD medium.

Keywords: 26S rDNA, ITS 1/5.8S rDNA/ITS2, wild flowers, yeasts, anti-gout xanthine oxidase inhibitor

*Corresponding author

Tel: +82-42-520-5388, Fax: +82-42-520-5388 E-mail: [email protected]

© 2013, The Korean Society for Microbiology and Biotechnology

(2)

국내야생화들의효모분포상을조사하고이들효모로부터 새로운고부가가치의생리활성물질을생산하고자먼저서울

(

구로동

⁾

을포함하는경기도지역과제주도송악산일대에서

서식하고있는진달래등의야생화들로부터효모를분리하 여동정하였고이들가운데항통풍성

xanthine oxidase

저 해물질을생산하는효모를선발하여생산조건을검토하였다

^.

재료 및 방법

효모의 분리

경기도지역

⁽

서울구로동

^,

평택

^,

성남

^,

용인

^,

안성

⁾

과제주

도송악산과성산일출봉일대에서

²⁰¹²

년봄에개화한꽃

(

화분

^{) 52}

점들을멸균튜브에채취하여

^,

멸균수에넣고

¹

시간 진탕시킨후일정액을

streptomycin (50

μ

^g/ml

과

ampicillin (50

μ

^g/ml)

을함유한

yeast extract-peptone dextrose

한천배 지에도말하여

³⁰

^o

^C

에서

⁴⁸

시간배양한후생육한효모들을 분리하였다

^.

효모의 동정

효모의 동정은

^{26S rDNA}

의

D1/D2 region

또는

^ITS1/

5.8S rDNA/ITS2 region

의염기서열상동성비교법을이용 하여다음과같이실시하였다

^.

먼저분리한효모들의

^gDNA

를

Genomic DNA prep kit for yeast (Sol Gent Co.)

로 추 출하여

^PCR

을위한주형

^DNA

로사용하였다

^.

분리한

^DNA

를

^{26S rDNA}

의

D1/D2 region primer

로

LN1 (5'GCA TAT CAA TAA GCG GAG GAA AAG 3')

과

^NL4(5'GGT CCG TGT TTC AAG ACG G 3')

또는

ITS1/5.8S rDNA/

ITS2 region primer

로

ITS1(5'TCC GTA GGT GAA CCT GCG G 3')

과

ITS4 (5'TCC TCC GCT TAT TGA TAT GC 3')

를이용하여

^PCR

하여증폭시킨후

1.5% agarose gel

을이 용한 전기영동으로 확인하고 다시 이

^PCR

산물을

^Gel extraction kit (Qiagen)

로정제하고이들의염기서열을분석 하였다

^.

염기서열의정렬은

^DNAstar

를사용하여

^Florence Corpet

로

Multiple sequence alignment

를하여염기서열을 정렬하였고

^{, NCBI}

의

^BLAST

의

^database

염기서열과의상 동성을조사하였다

^[20].

Xanthine oxidase 저해활성 측정

Xanthine oxidase

저해활성은먼저효모를

³⁰

^o

^C

로

²

일간

YEPD

배지에서배양한후배양상등액과무세포추출물로

구분하여각각동결건조시킨다음

0.1 M potassium phosphate buffer (pH7.5)

에

^{10 mg/ml}

로용해하였다

^.

이들시료

¹⁰⁰

μ

^l

를

1 mM xanthine

용액

²⁰⁰

μ

^l

와

xanthine oxidase (0.1 U/ml) 100

μ

^l

에혼합하고

0.1 M potassium phosphate buffer (pH7.5) 600

μ

^l

첨가하여

^{1 ml}

로정용한후

³⁷

^o

^C

에서

5

분간각각반응시켰다

. 1 N HCl 200

μ

^l

를가하여반응을정 지시킨후

12,000 rpm

으로

¹⁰

분간원심분리하여단백질을 제거한다음생성된

uric acid

함량을분광분석기

^(UV-1601, Shimadzu)

로

^{292 nm}

에서흡광도를측정하여정량한후아 래와같이배양상등액과무세포추출물의

xanthine oxidase

저해활성을계산하였고

^{[6] 3}

반복의평균값으로표시하였다

^.

xanthine oxidase

저해활성

^{(%) =}

[1

−

^{A(

시료구

⁾

−

^B(

시료구

^Blank)/C(

대조구

^)}]

×

¹⁰⁰

결과 및 고찰

경기도 야생화들로부터 효모의 분리 및 동정

서울구로동과평택

^,

안성

^,

용인

^,

성남등지의야생화들로 부터효모들을분리하여이들의

^{26S rDNA}

의

D1/D2, ITS1/

5.8S rDNA/ ITS2

염기서열분석을통한분자생물학적유연 관계비교분석방법으로동정한결과는

^{Table 1}

과같다

^.

각기다른

³¹

점의야생화들로부터

²⁶

종의효모들을

⁵⁴

균 주분리하였다

^.

이들지역의야생화들로부터

Pseudozyma rugulosa

를포함하는

Pseudozyma sp.

가

¹³

균주분리되어이 들지역에서는이균주들이우점균이었고

Cryptococcus aureus

균주를 포함한

Crytococcus sp.

가

⁷

균주

, Rhodosporidium sp.

와

Rhodotonula sp.

가각각

⁴

균주

^{, 5}

균주가분리되었다

^.

이밖에도

Sporobolomyces sp.

과

Metschnikowia sp.

등이 분리되었다

^.

또한일반적으로전통발효식품등에서유용균 주로알려진

Saccharomyces cerevisiae

종이

⁴

균주분리되었 다

^.

이들중우점균들에대한최근연구로

^Morita

등

^[22]

은 당지질성생계면활성제인

mannosylerythritol(MEL)

생산능 력이우수한

Pseudozyma rugulosa

를분리하여동정한후

MEL

최적생산조건을보고하였고

, Pseudozyma fusiformata

를 포함하는몇종의

Pseudozyma sp.

들역시

^MEL

생성능이우 수함을보고하였다

^[23].

또한

^{, Cao}

등

^[1]

은해양효모로알려 진

Cryptococcus aureus HYA

가갖고있는

^insulin

분해효소 유전자인

^INU1

을

Pichia pastoris X-33

에클로닝시켜대량 발현시킨 후 이들의 특성을 보고하였고

^{, Gao}

등

^[4]

은

Cryptococcus aureus G7

이

Jerusalem artichoke

추출물에 서단세포단백질을많이생산하였음을보고하였다

^.

제주도 야생화들로부터 효모의 분리 및 동정

제주도송악산과성산일출봉지역산과밭등지의

²¹

점 의꽃들로부터효모들을분리하여동정한결과모두

¹⁴

종에 속하는 효모들을

³²

균주 분리하였다

(Table 2).

이들 중

Sporobolomyces ruberrimus

가

⁶

균주

, Pseudozyma aphidis

4

균주를포함한

Pseudozyma sp.

가

⁸

균주

, Candida sp.

가

⁵

균주등으로우점균을이루고있었다

^{. Fell}

등

^[3]

은

^{D1, D2}

(3)

Table 1. Yeasts species from wild flowers in Gyonggi-do including Guro-dong, Seoul.

No. Putative species Isolated

No.

Related Genbank sequence

Identity (%)

Collected area

1 Cryptococcus aureus 225-S-2 EU304246.1 598/600 (99%)

Seoul Guro-dong

2 Cryptococcus laurentii 222-S-2 HQ327003.1 578/588 (98%)

3 Lachancea thermotolerans 225-S-1 CU928180.1 578/581 (99%)

227-S-2 CU928180.1 577/581 (99%)

4 Pseudozyma hubeiensis 222-S-1 AB566327.1 605/607 (99%)

228-S-1 AB566327.1 609/615 (99%)

5 Rhodotorula glutinis 222-S-3 JQ917424.1 570/576 (99%)

6 Sporobolomyces carnicolor 228-S-2 JQ964222.1 497/503 (99%)

7 Sporobolomyces carnicolor 226-S-1 JN940713.1 604/604 (100%)

8 Sporobolomyces phaffii 224-S-2 AY070011.1 600/601 (99%)

223-S-3 AY070011.1 601/601 (100%)

9 Candida sp. 277-GG1-2 AY740171.1 647/648 (99%)

Pyeongtaek city

10 Cryptococcus aureus 282-GG1-3 EU304246.1 600/602 (99%)

11 Cryptococcus flavus 274-GG1-1 FJ743627.1 638/640 (99%)

12 Metschnikowia koreensis 277-GG1-1 AF296438.1 532/532 (100%)

13 Pseudozyma rugulosa 269-GG1-1 JN940523.1 604/607 (99%)

269-GG1-2 JN940523.1 604/607 (99%) 269-GG1-3 JN940523.1 613/618 (99%) 270-GG1-1 JN940523.1 605/607 (99%) 271-GG1-1 JN940523.1 648/648 (100%) 14 Rhodosporidium fluviale 281-GG1-1 FJ515261.1 612/616 (99%)

15 Rhodotorula graminis 282-GG1-1 EU563930.1 617/617 (100%)

16 Saccharomyces cerevisiae 272-GG1-1 JX103177.1 607/608 (99%) 274-GG1-2 JX103177.1 608/608 (100%) 17 Sporidiobolus pararoseus 270-GG1-2 FJ755245.1 574/577 (99%) 18

Hanseniaspora uvarum

319-GS-1 AM160628.1 614/615 (99%)

Seongnam city 325-GS-1 AM160628.1 615/615 (100%)

326-GS-1 AM160628.1 615/615 (100%) 327-GS-1 AM160628.1 613/615 (99%)

19 Metschnikowia reukaufii 324-GS-1 JX067756.1 545/557 (98%)

20 Metschnikowia viticola 326-GS-2 JN544019.1 443/529 (84%)

21 Pichia guilliermondii 321-GS-1 FJ432597.1 613/613 (100%)

22 Pseudozyma rugulosa 323-GS-1 JN940523.1 648/648 (100%)

23 Saccharomyces cerevisiae 322-GS-2 JX103177.1 607/608 (99%) 24 Sporidiobolus pararoseus 323-GS-2 JQ219312.1 575/576 (99%) 25 Cryptococcus flavescens 330-GY-1 FJ743610.1 640/640 (100%)

Yongin city

26 Pichia fermentans 331-GY-2 DQ377652.1 598/599 (99%)

27 Pseudozyma rugulosa 328-GY-3 JN940523.1 644/644 (100%)

329-GY-2 JN940523.1 644/644 (100%) 330-GY-2 JN940523.1 647/648 (99%) 28 Rhodosporidium paludigenum 328-GY-1 HQ670686.1 617/617 (100%)

329-GY-1 FJ794995.1 617/617 (100%

29 Rhodotorula mucilaginosa 331-GY-4 HQ871906.1 576/577 (99%)

30 Rhodotorula sp. 332-GY-2 EU678938.1 622/627 (99%)

31 Saccharomyces cerevisiae 328-GY-2 JX103177.1 605/608 (99%) 32 Sporisorium loudetiae-pedicellatae 332-GY-3 AY740106.1 645/649 (99%)

33 Candida sp. 333-GW-1 AY833088.1 647/648 (99%)

Anseong city 334-GW-1 AY833088.1 647/648 (99%)

34 Cryptococcus sp. 339-GW-1 FM178286.1 606/607 (99%)

337-GW-1 GQ181170.1 633/639 (99%)

35 Pseudozyma rugulosa 335-GW-1 JN940523.1 604/607 (99%)

337-GW-2 JN940523.1 604/607 (99%)

36 Rhodosporidium fluviale 340-GW-2 FJ515261.1 612/616 (99%)

37 Rhodotorula mucilaginosa 340-GW-1 JX310560.1 607/613 (99%)

(4)

와

^ITS

영역에있는

^rDNA

염기조성과형태적특성등을토 대로

Sporobolomyces ruberrimus

를분리

^,

동정하여보고하 였고

^{, Razavi}

등

^[26]

은

Sporobolomyces ruberrimus

로부터 글리세롤과암모니움아세테이트를사용하여카로티노이드 와 지방산 등을 생산한 연구결과를 보고하였다

^.

또한

Pseudozyma aphidis

균에대하여최근

^Parahym

등

^[24]

은 폐감염균종으로

^{, Chen}

등

^[2]

은다리감염균종으로각각보 고하였다

^.

한편

^,

필자등

^[20]

은대전지역공단주위와야산의야생화 로부터희귀성균주들인

Metschnikowia sp.

와

Starmerella sp.

들이

Sporobolomyces rubberimus, Cryptococcus magnus

등을분리

^,

보고하였는데위와같이동일한야생화라도경기 도

^,

대전

^,

제주도등지역간의기후적차이와이로인한개 화시기차이등에의해일부다른효모들이분포하고있는

것으로사료된다

^.

항통풍성 xanthine oxidase 저해물질 생산 균주의 선발 경기도와제주도야생화들로부터분리

^,

동정한

⁸⁶

균주들 에대한배양상등액과무세포추출물들의

xanthine oxidase

저해활성을측정한결과는

^{Table 3}

과같다

^.

먼저

⁸⁶

균주중

26

균주들의배양상등액에서

xanthine oxidase

저해활성을 보였고

^,

이들중특히

Rhodosporidium paludigenum 328- GY-1

의배양상등액이

^18.1%

로가장높은활성을보였다

^.

무세포추출물의경우모두

²⁴

효모균주들에서저해활성 을보였고특히

Pseudozyma hubeiensis 228-S-1

의무세포 추출물이

^19.6%

로가장높아최종적으로항통풍성

^xanthine oxidase

저해물질생산균주로선발하였다

^.

항통풍성

xanthine oxidase

저해물질생산균주로선발된

Table 2. Yeasts species from wild flowers in Songak Mt. and Sungsan-ilchulbong of Jeju island, Korea.

No. Putative species Isolated

No.

Related Genbank

sequence Identity (%)

1 Candida parapsilosis 78-J-1 HE605209.1 612/613(99)

2 Candida sp. 80-J-3 DQ655679.1 588/592 (99)

90-J-1 HM222574.1 530/531 (99)

78-J-2 HM222578.1 529/530 (99)

67-J-1 AY740167.1 404/424 (95)

3 Cryptococcus aureus 86-J-3 HQ832810.1 528/529 (99)

4 Debaryomyces hansenii 80-J-2 JQ026331.1 321/327 (98)

89-J-2 JQ026331.1 321/327 (98)

5 Metschnikowia reukaufii 67-J-3 FJ455114.1 550/554(99)

6 Metschnikowia pulcherrima 66-J-1 AY235809.1 332/338 (99)

7 Nakazawaea holstii 63-J-1 AB449811.1 628/643 (97)

83-J-1 AB449811.1 628/643 (97)

8 Pichia guilliermondii 89-J-1 EU568971.1 603/607 (99)

9 Pseudozyma aphidis 66-J-2 HQ832804.1 440/440 (100)

77-J-1 JN942666.1 439/441 (99)

64-J-1 N942666.1 439/441 (99)

94-J-3 JN942666.1 439/441 (99)

10 Pseudozyma rugulosa 84-J-1 JN942670.1 726/737 (98)

86-J-2 AF294697.1 718/742 (96)

90-J-2 AF294697.1 726/728 (99)

11 Pseudozyma sp. 71-J-1 AM160637.1 441/442 (99)

12 Rhodosporidium paludigenum 86-J-1 HQ670676.1 605/607 (99)

89-J-3 HQ670676.1 607/607 (100)

92-J-2 HQ670676.1 605/607 (99)

94-J-2 HQ670676.1 607/607 (100)

13 Sporobolomyces ruberrimus 63-J-2 AY015439.1 595/598 (99)

65-J-1 AY015439.1 588/602 (97)

70-J-1 AY015439.1 588/602 (97)

87-J-1 AY015439.1 588/602 (97)

94-J-1 AY015439.1 588/602 (97)

95-J-2 AY015439.1 588/602 (97)

14 Starmerella bombicola 80-J-1 HQ111046.1 504/506(99)

(5)

Table 3. Xanthine oxidase (XOD) inhibitory activities of the isolated yeasts. (Unit: %) Yeast strains Extracelluar XOD inhibitory activity Intracelluar XOD inhibitory activity

1 Candida parapsilosis 78-J-1 4.4(±0.5) n.d

^a

2 Candida sp. 277-GG1-2 n.d n.d

333-GW-1 n.d n.d

334-GW-1 n.d n.d

67-J-1 n.d n.d

78-J-2 10.4(±0.6) n.d

80-J-3 5.6(±0.8) n.d

90-J-1 n.d 4.7(±0.7)

3 Cryptococcus aureus 86-J-3 n.d n.d

225-S-2 n.d n.d

282-GG1-3 n.d n.d

4 Cryptococcus flavescens 330-GY-1 n.d n.d

5 Cryptococcus flavus 274-GG1-1 7.3(±0.2) n.d

6 Cryptococcus laurentii 222-S-2 6.2(±0.6) 8.7(±0.5)

7 Cryptococcus sp. 337-GW-1 7.8(±0.9) n.d

339-GW-1 n.d n.d

8 Debaryomyces hansenii 80-J-2 n.d n.d

89-J-2 n.d n.d

9 Hanseniaspora uvarum 319-GS-1 5.2(±0.2) n.d

325-GS-1 n.d n.d

326-GS-1 12.5(±0.3) n.d

327-GS-1 n.d n.d

10 Lachancea thermotolerans 225-S-1 13.4(±0.4) 9.7(±0.8)

227-S-2 9.9(±0.5) 8.5(±0.4)

11 Metschnikowia koreensis 277-GG1-1 16.8(±0.9) n.d(±0.6)

12 Metschnikowia pulcherrima 66-J-1 6.8(±0.3) n.d

13 Metschnikowia reukaufii 67-J-3 n.d n.d

324-GS-1 n.d n.d

14 Metschnikowia viticola 326-GS-2 13.1(±0.5) 17.8(±0.9)

15 Nakazawaea holstii 63-J-1 n.d n.d

83-J-1 n.d n.d

16 Pichia fermentans 331-GY-2 n.d n.d

17 Pichia guilliermondii 89-J-1 n.d n.d

321-GS-1 n.d n.d

18 Pseudozyma aphidis 64-J-1 n.d n.d

66-J-2 9.4(±0.5) n.d

77-J-1 15.2(±0.6) n.d

94-J-3 n.d

19 Pseudozyma hubeiensis 222-S-1 n.d n.d

228-S-1 n.d 19.6(±0.8)

20 Pseudozyma rugulosa 269-GG1-1 n.d 4.1(±0.3)

269-GG1-2 n.d n.d

269-GG1-3 n.d n.d

270-GG1-1 n.d 7.5(±0.5)

271-GG1-1 n.d 12.4(±0.7)

323-GS-1 5.1(±0.5) 17.8(±0.5)

328-GY-3 n.d n.d

329-GY-2 n.d n.d

330-GY-2 n.d n.d

335-GW-1 12.9(±0.8) 15.8(±0.6)

337-GW-2 n.d n.d

84-J-1 n.d n.d

86-J-2 1.8(±0.9) 9.5(±0.9)

90-J-2 n.d n.d

(6)

Pseudozyma hubeiensis 228-S-1

은필자등이국내미기록 효모로보고

^[5]

한균주로서국내에서는이균의특성이나산 업적응용성에관한연구는실시되지않았고다만

^Takahashi

등

^[27]

이

Pseudozyma hubeiensis

의대두유배양물에함유된

mannosylerythritol

과이들의유도체가항산화활성과

^SOD

활성이우수하였음을보고하였을뿐이다

^.

Xanthine oxidase 저해물질 생산조건

Pseudozyma hubeiensis 228-S-1

의

xanthine oxidase

저 해물질생산에미치는배양시간의영향을조사한결과는

^Fig.

1

과같다

. Pseudozyma hubeiensis 228-S-1

을

^YEPD

배지에 접종하여

³⁰

^o

^C

에서배양하였을때생육은

⁴⁸

시간에최고를

보였으나항통풍성

^XOD

저해물질은대수기말기인

³⁶

시간

배양한무세포추출물에서제일높았다

^. Table 3. Continued.

Yeast strains Extracelluar XOD inhibitory activity Intracelluar XOD inhibitory activity

21 Pseudozyma sp. 71-J-1 n.d n.d

22 Rhodosporidium fluviale 281-GG1-1 7.4(±0.5) n.d

340-GW-2 n.d 10.2(±0.4)

23 Rhodosporidium paludigenum 86-J-1 n.d n.d

89-J-3 16.5(±0.6) n.d

92-J-2 n.d 3.8(±0.9)

94-J-2 n.d n.d

328-GY-1 18.1(±0.5) n.d

329-GY-1 n.d 7.3(±0.9)

24 Rhodotorula glutinis 222-S-3 6.1(±0.5) n.d

282-GG1-1 9.1(±0.6) n.d

25 Rhodotorula mucilaginosa 331-GY-4 n.d 12.0(±0.7)

340-GW-1 n.d n.d

26 Rhodotorula sp. 332-GY-2 4.4(±0.3) 3.2(±0.9)

27 Saccharomyces cerevisiae 272-GG1-1 n.d 1.9(±0.2)

274-GG1-2 n.d 2.4(±0.1)

322-GS-2 n.d 2.7(±0.6)

328-GY-2 n.d 1.9(±0.9)

28 Sporidiobolus pararoseus 270-GG1-2 11.8(±0.9) 7.7(±0.8)

323-GS-2 n.d n.d

29 Sporisorium loudetiae-pedicel-

latae 332-GY-3 n.d 18.6(±0.5)

30 Sporobolomyces carnicolor 226-S-1 n.d n.d

228-S-2 9.8(±0.9) n.d

31 Sporobolomyces phaffii 223-S-3 n.d n.d

224-S-2 n.d n.d

32 Sporobolomyces ruberrimus 63-J-2 n.d

65-J-1 n.d n.d

70-J-1 n.d n.d

87-J-1 n.d n.d

94-J-1 n.d n.d

95-J-2 n.d n.d

33 Starmerella bombicola 80-J-1 n.d 17.8(±0.9)

a

n.d; not detected.

Fig. 1. Effect of cultural time on the xathine oxidse (XOD)

inhibitory activity of Pseudozyma hubeiensis 228-S-1.

(7)

항통풍성

^XOD

저해활성물질에관한주요연구로

^Song

등

^[25]

은

Phellinus linteus

버섯자실체의

^70%

에탄올추출 물이

^XOD

저해활성을보였다고보고하였고서등

^[19]

은청 미래덩굴잎발효차의에탄올추출물에서

^,

김등

^[13]

은주박

(

술지게미

⁾

의열수와에탄올추출물에서

^XOD

저해활성이있 었음을보고하였다

^.

그러나아직까지이들의생산과산업적 이용및물질특성에관한연구는실시되지않았고따라서 최근증가하고있는통풍질환을예방하기위해배양이용 이하고단시간에대량배양이가능한본연구의선발균주를 이용한항통풍성건강소재개발에관한연구가현재진행되 고있다

^.

요 약

국내야생화들로부터효모를분리하여이들을고부가가치 의의약이나건강식품산업에응용하기위한연구의일환으 로경기도와제주도일부지역의야생화들을채집하여효모

들을 분리

^,

동정하고 이들 가운데 항통풍성

^xanthine

oxidase

저해물질생산균주를선발한후생산조건을검토

하였다

^.

서울구로동과일부경기도지역야생화

³¹

점에서

26

종

^{, 54}

균주의 효모들을 분리

^,

동정하였고 이들 중

Pseudozyma sp.

가

¹³

종으로가장많이분리되었다

^.

제주도 송악산과성산일출봉지역의야생화

²¹

점에서모두

¹⁴

종

^, 32

균주의 효모들을 분리

^,

동정하였고

^,

이들 균 중에서

Sporobolomyces ruberrimus

가

⁶

종으로가장 많이분리되 었다

^.

86

효모균주들을

^YEPD

배지에서

²

일간배양하여각각의 배양상등액과무세포추출물을제조한후이들의항통풍성

Xanthine oxidase

저해활성을 측정한 결과

, Pseudozyma hubeiensis 228-S-1

의무세포추출물이

^19.5%

의가장높은

Xanthine oxidase

저해활성을 보였다

^.

또한

Pseudozyma hubeiensis 228-S-1

을

^YEPD

배지에 접종하여

³⁰

^o

^C

에서

36

시간배양하였을때

^,

세포내항통풍성

xanthine oxidase

저해물질을가장많이생산하였다

^.

Acknowledgments

This study was funded by the project on survey and excavation of Korean indigenous species of NIBR under the Ministry of Envi- ronment, Republic of Korea.

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